Leading chaser Shishkin suffered a fatal injury in his box on Sunday evening, trainer Nicky Henderson has reported.
The 10-year-old was on course to run in next Wednesday's Punchestown Gold Cup but was found cast in his box on Sunday evening and fractured a leg. He could not be saved.
Henderson reported on social media: "Tragically we have lost Shishkin this evening after he got cast in his stable and fractured his hind leg. He was due to leave for Punchestown tomorrow but sadly this is now not to be.
"He was an absolute superstar and his CV is testament to that - except the King George is not in it though we believe it should have been.
"The Supreme Novice Hurdle and the Arkle were his Cheltenham highlights, but the battle with Energumene in the Clarence House at Ascot was probably his greatest.
"This is a very sad night, He was our star and we will never forget him. Thank you Shishkin for the wonderful memories."
By Sholokhov, Shishkin started his career in Ireland, winning a point-to-point for Virginia Considine in November 2018. He was sold for £170,000 to Highflyer Bloodstock who were acting on behalf of owners Joe and Marie Donnelly.
Shishkin was a 6/1 shot when he won the 2020 Supreme Novices' Hurdle by a head from Abacadabras.
He was sent chasing the following season and won all five of his starts, including the Arkle at Cheltenham and the Maghull Novices' Chase at Aintree.
Moved up into open company, he took on and defeated Energumene in an unforgettable clash at Ascot in January 2022 but the race appeared to leave a mark on him and he was pulled up in the Champion Chase two months later when an odds-on favourite.
Since then his form has been in-and-out but he looked as good as ever at Aintree in April 2023 when stepped up to three miles, winning the Aintree Bowl Chase in fine style.
Last Christmas he looked to have the King George VI Chase at his mercy, holding the lead when stumbling and unseating jockey Nico de Boinville after the second last fence. The race was won by Hewick.
Shishkin bounced back to form in a Grade 2 chase at Newbury in February and was considered a serious contender for the Gold Cup before being withdrawn a couple of days before the race due to fears that the Henderson horses were off-form.
At Aintree earlier this month Shishkin finished a moderate fourth behind Gerri Colombe and was reported to be in good form ahead of Punchestown where he was due to take on Galopin Des Champs.